Szczerbakowa A, Maciejewska U, Zimnoch-Guzowska E, Wielgat B
Institute of Biochemistry and Biophysics PAS, Pawińskiego 5A, 02-106 Warsaw, Poland.
Plant Cell Rep. 2003 Feb;21(6):577-84. doi: 10.1007/s00299-002-0555-8. Epub 2002 Dec 12.
Somatic hybrids between the cultivated potato diploid hybrid clone, ZEL-1136, and hexaploid non-tuber-bearing wild species Solanum nigrum L. exhibiting resistance to Phytophthora infestans were regenerated after PEG-mediated fusion of mesophyll protoplasts. The objective was to transfer the late-blight resistance genes from the wild species into plants of the cultivated potato clone. From a total of 59 regenerants, 40 clones survived and have been maintained in vitro on hormone-free MS/2 medium. Thirty-two somatic hybrids were identified by their intermediate morphology (leaves of nigrum type and flowers of tuberosum type) and verified by flow cytometry and random amplified polymorphic DNA (RAPD) patterns. The RAPD analysis of nuclear DNA confirmed the hybrid nature of 29 clones. Flow cytometry revealed a wide range of ploidy in the generated hybrids, from nearly the tetra- to decaploid level. Most of the hybrid clones were stable in vitro, grew vigorously in soil, and set flowers and parthenocarpic berries. However, all of the flowering hybrids were male-sterile. Nine hybrid clones produced tuber-like structures in soil. The most vigorous flowering somatic hybrids were selected for assessment of the late-blight resistance.
在通过聚乙二醇介导的叶肉原生质体融合后,获得了栽培马铃薯二倍体杂交克隆ZEL - 1136与对致病疫霉具有抗性的六倍体非块茎野生种龙葵之间的体细胞杂种。目的是将野生种的晚疫病抗性基因转移到栽培马铃薯克隆的植株中。在总共59个再生植株中,40个克隆存活下来,并在无激素的MS/2培养基上进行离体保存。通过其中间形态(龙葵型叶片和马铃薯型花朵)鉴定出32个体细胞杂种,并通过流式细胞术和随机扩增多态性DNA(RAPD)图谱进行了验证。对核DNA的RAPD分析证实了29个克隆的杂种性质。流式细胞术显示所产生的杂种中存在广泛的倍性范围,从近四倍体到十倍体水平。大多数杂种克隆在离体条件下稳定,在土壤中生长旺盛,并能开花和结单性结实的浆果。然而,所有开花的杂种都是雄性不育的。9个杂种克隆在土壤中产生了类似块茎的结构。选择最旺盛开花的体细胞杂种来评估晚疫病抗性。